1. Remove media from around and on top of gel, wash 3 x 5 minute incubations with PBS.
2. Remove gel from insert and add to homogeniser tube
3. Add 400uL of pronase solution (10mg/mL stock) (Sigma)
4. Triturate the gel
5. Incubate in water bath at 37°C for 5 minutes, with agitation every minute
6. Add buffer RLT from RNeasy mini kit (350μl for <5×106 cells)
7. Homogenise with an automated homogeniser (our lab uses IKA RW16 basic, Sigma)
8. Continue with RNeasy protocol